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WO1992003548A1 - Proteine zp3 de la zona pellucida humaine - Google Patents

Proteine zp3 de la zona pellucida humaine Download PDF

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Publication number
WO1992003548A1
WO1992003548A1 PCT/EP1991/001538 EP9101538W WO9203548A1 WO 1992003548 A1 WO1992003548 A1 WO 1992003548A1 EP 9101538 W EP9101538 W EP 9101538W WO 9203548 A1 WO9203548 A1 WO 9203548A1
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Prior art keywords
human
polypeptide
polypeptide according
antibodies
acid sequence
Prior art date
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Ceased
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PCT/EP1991/001538
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English (en)
Inventor
Marcel Van Duin
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Akzo NV
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Akzo NV
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Publication date
Application filed by Akzo NV filed Critical Akzo NV
Priority to FI930897A priority Critical patent/FI930897A7/fi
Priority to KR1019930700637A priority patent/KR930702522A/ko
Publication of WO1992003548A1 publication Critical patent/WO1992003548A1/fr
Priority to NO93930718A priority patent/NO930718L/no
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues

Definitions

  • the invention relates to a polypeptide or a functional derivative thereof having human ZP3 activity or having, at least in part, human ZP3 antigenicity.
  • ZP zona pellucida
  • the ZP is an extracellular matrix which comprises three glycoproteins, designated ZP1, ZP2 and ZP3, of which ZP3 has been identified as the sperm receptor (reviewed in Wassarman, Development 108, 1-17; 1990).
  • ZP3 is an important candidate target antigen in experimental strategies aimed at the development of immuno-contraception (Henderson et al., J. Reprod. Fert. 83, 325-343; 1988 and refs.
  • ZP3 for contraception was emphasized by studies that revealed a long term infertility following vaccination of female mice with an oligopeptide derived from the murine ZP3 amino acid sequence (Millar et al., Science 246, 935-938; 1989).
  • mouse polypeptides are not suitable since the ZP3 sperm receptor is species specific. Moreover, polypeptides from non-human origin may lead to unwanted immune responses in man. Therefore, development of a safe and efficient contraceptive vaccine based on (part of) the human sperm receptor requires information on and availability of the human ZP3 polypeptide. The isolation of sufficient amounts of human ZP3 polypeptide from female gametes is of course not possible. However, we have succeeded in identifying a human gene and elucidating its sequence. This opens the possibility to produce ZP3 polypeptides and/or ZP3 polypeptide fragments either by recombinant DNA technology or solid phase polypeptide synthesis.
  • the invention therefore comprises a novel DNA-molecule coding for at least a part of human ZP3 polypeptide.
  • Said DNA-molecule comprises at least a part of the sequence shown in Fig. 2.
  • the invention also comprises a polypeptide coded for by at least part of the above-mentioned DNA-molecule.
  • Said polypeptide may comprise at least a part of the amino acid sequence shown in Fig. 2.
  • Functional derivatives are meant to include polypeptides in which one or more of the amino acids have been replaced with a chemically comparable one.
  • the crux is of course that they should still show human ZP3 antigenicity.
  • polypeptides according to the invention can be produced either synthetically or by recombinant DNA technology. Methods for producing synthetic polypeptides are well known in the art and do not need any further elaboration.
  • polypeptides by recombinant DNA techniques is a general method which is known, but which has a lot of possibilities all leading to somewhat different results.
  • the polypeptide to be expressed is coded for by a DNA sequence or more accurately by a nucleic acid sequence.
  • This nucleic acid sequence must be transcribed (optionally) and translated to the wanted polypeptide.
  • the nucleic acid sequence is normally cloned into a vector with which a host cell is transformed.
  • the vector can be either self replicating or it may integrate into the DNA of the host.
  • Different host cells lead to different polypeptides. Prokaryotes do not possess the organelles necessary for glycosylation. The polypeptides produced by them will be without carbohydrate side chains. Eukaryotes do have the glycosylation machinery, but yeast cells will give a different glycosylation pattern than mammalian cells.
  • polypeptides according to the invention is an expression system which gives the most "natural" glycosylation pattern. Towards this end mammalian cells are most preferred.
  • epitopes on the human ZP3 polypeptide which may have contraceptive potential. These epitopes comprise at least a part of one of the following sequences:
  • Small polypeptides like these are more easily produced synthetically. They can also be linked together with the same or different "epitopes" to form a larger more antigenic polypeptide.
  • the vaccination may be either a passive or an active immunization.
  • a polypeptide according to the invention is administered to a female (possibly with an adjuvant).
  • the administration will give rise to an immune response by the female.
  • Antibodies will be produced which recognize human ZP3 on the ovum. These antibodies will specifically bind to the sperm receptor binding site so that spermatozoa cannot bind, or otherwise the antibodies will prevent this binding through steric hindrance.
  • Passive immunization basically works the same way. Instead of the antigen or its mimicry the antibodies against it are directly administered. Thus, antibodies have to be raised against the polypeptides according to the invention.
  • the B-lymphocytes of the mammal are harvested after a suitable period of time and immortalized through fusion or transformation. These methods are well known in the art.
  • Antibodies can be isolated from the culture of the immortalized lymphocytes.
  • Human antibodies can be produced by in vitro stimulation of isolated B- lymphocytes, or they can be isolated from (immortalized) B-lymphocytes which have been harvested from a female immunized with at least one polypeptide according to the invention.
  • Another object of the invention is the use of ZP3 polypeptide and antibodies directed to it in diagnostic test kits.
  • a human ovary gtlO cDNA library was purchased from Stratagene.
  • Oligonucleotides were prepared on an Applied Biosystems 381 A DNA synthesizer and directly used for cloning purposes.
  • Oligopeptides were produced by solid phase peptide synthesis using procedures described by Fields and Noble (Int. J. Pept. Prot. Res. 35, 160-214;
  • Total human zona pellucida Salt stored human zonae pellucidae were heat solubilized and mixed with freunds adjuvants for immunization of rabbits. Antiseraum titers were assayed on ELISA plates coated with porcine ZP-proteins. ⁇ Gal-ZP3 fusion protein. The fusion protein was partially purified from sonicated bacteria based on its insolubility and separated by SDS polyacrylamide gelelectrophoresis (SDS-PAGE). By electro-blotting the proteins were transferred to a nitrocellulose membrane. The region carrying the hybrid protein was excised and dissolved in DMSO. This was mixed in a 1:1 ratio with Freunds adjuvants and used for immunization of rabbits.
  • SDS-PAGE SDS polyacrylamide gelelectrophoresis
  • CHO produced ZP3.
  • a similar procedure was followed for the ZP3 protein produced by Chinese hamster ovary (CHO) cells. Concentrated culture medium was separated by SDS-PAGE. The 40-60 kiloDalton region was excised and used for immunization of mice.
  • Oligopeptides Synthesized peptides were physically linked to keyhole limpet hemocyanin (KLH) and injected in rabbits. As a control rabbits were immunized with KLH only. Sera were screened on ELISA microtitre plates coated with peptide. Human egg fluorescence assay.
  • KLH keyhole limpet hemocyanin
  • Salt stored unfertilized human eggs (after 48 hour incubation with sperm in an IVF program) were incubated with antisera (1:50 diluted in buffer A [PBS + 5% BSA]). After three washes eggs were incubated with a second antibody (swine a i rabbit or mouse conjugated to FITC, 1:100 diluted in buffer A) and incubated for 1 hour at 37 °C. Following three additional wash steps fluorescence was measured with a Nikon microscope and exposure analyser. Non-stained (negative) control eggs are dark, with a long exposure time, and positively stained zonae pellucidae have shorter exposure times.
  • a 135 bp mouse ZP3 probe was constructed by assembling 8 synthetic oligonucleotides (27-51 mers). This fragment, comprising part of exon 5 and 6 of the murine gene (position 771 to 909 in Ringuette et al., Developmental Biology 127, 287-295; 1988) was provided with unique 5' BamHI and 3' Hindlll restriction sites and subcloned in pGEM3 (Promega). Subsequently its nucleotide sequence was verified.
  • Fig. 1 Restriction fragments from clones II and Dl were subcloned in pGEM vectors and M13mpl8/19 and used for genomic characterization of the human gene. This involved localization of the exons by Southern blot analysis with ⁇ 2 P-labeled oligonucleotides from the mouse ZP3 sequence followed by sequence analysis. As is demonstrated in Fig.
  • the human gene consists of 8 exons spread over approximately 20 kb of genomic DNA. All identified exons are flanked by consensus splice donor and splice acceptor signals (not shown). From sequence analysis of all exons the complete coding sequence for the ZP3 gene could be deduced (presented in Fig. 2).
  • the human ZP3 gene encodes a polypeptide of 372 amino acids with a calculated molecular weight of 41437 Dalton.
  • a human ovary cDNA library (gtlO) was screened with 3 P-labeled fragments from human ZP3 exon 1, 5 and 7. This yielded several partial cDNA's that contained only the 3' half of the ZP3 coding sequence ( exon 5 Sequence analysis of 3 independent cDNA clones confirmed the previously determined genomic ZP3 sequence except for one residue in exon 7 (see Fig. 2). At nucleotide position 1064 a G and C residue was found in genomic and cDNA respectively. In the encoded amino acid sequence this yields an arginine or a threonine residue respectively at position 345. This sequence difference probably represents a polymorphism in the human ZP3 gene and polypeptide.
  • the resulting 2.7 kb 'minigene' contains a truncated intron between exon 2 and 3 and the natural introns between ZP3 exons 3 and 4 and exon 4 and 5.
  • the integrity of the polypeptide encoding nucleotide information of this ZP3 construct was completely verified by sequence analysis.
  • the ZP3 ' minigene' was subsequently inserted in a mammalian expression vector in which the ZP3 gene is driven by the strong SV40 early promoter.
  • this vector harbored ⁇ -Globin splicing and SV40 poly-adenylation signals for correct RNA processing of the expressed gene and the selectable marker gene aminoglycosylphosphotransferase (Colbere-Garapin et al., J. Mol. Biol. 150, 1-14, 1981) which allows for isolation of stable transformants by selection for G418 resistance.
  • CHO cells were transfected with the ZP3 expression construct using the calcium-phosphate precipitation technique (Graham and van der Eb, Virology 52, 456-467, 1973; Wigler et al, Proc. Natl. Acad. Sci USA 76,1373-1376, 1979).
  • Mass populations of G418 resistant transformants (representing 300-500 independent clones) were analysed for expression of the recombinant ZP3 gene.
  • Northern blot analysis of total RNA from pooled transformants with a 32 P-labeled ZP3 probe reveals a relatively high level of expression of the transfected ZP3 gene (as compared to the highly expressed actin gene, not shown).
  • minigene RNA is correctly processed to a mRNA which is slightly larger then the natural transcript found in human ovary RNA (see Fig. 4). This size difference is most likely due to flanking 5' and 3' sequences present in the expression vector.
  • ZP3 cDNA For expression in E.coli first a full lenght ZP3 cDNA was constructed using a cDNA fragment covering exon 1 to 5 isolated by PCR from CHO transformants and the partial cDNA clone (harboring exon 5 to 8) isolated from a human ovary cDNA library. Four ZP3 cDNA fragments were cloned in frame to the E.coli LacZ gene in pEX-plasmids (Biores) allowing the production of a ⁇ -galactosidase-ZP3 fusion polypeptide as reported by Stanley and Luzio, EMBO J. 3, 1429-1434, 1984).
  • fusion polypeptides in E.coli was examined by SDS polyacrylamide gelelectrophoresis (SDS-PAGE) and western blot analysis. The results of these experiments are presented in Fig. 5. Following heat shock induction, all four LacZ-ZP3 constructs yielded fusion polypeptides , although to a much lesser extend then the LacZ gene. The molecular weight of the hybrid polypeptides, as observed on coomassie stained gels (see Fig. 5A), were in agreement with the expressed ZP3 parts. This was further confirmed by western blot analysis with anti- ⁇ -galactosidase antibodies (not shown) and a rabbit antiserum raised against ZP3 amino acids 341 to 360 coupled to KLH (Fig. 5B).
  • fusion polypeptides carrying this region i.e. ZP3-A and ZP3- D, were recognized by the antiserum.
  • An antiserum against KLH yielded a result similar to the background staining in Fig. 5B (not shown).
  • BGal-ZP3 fusion polypeptides were recognized by antisera raised against total human zonae pellucidae (not shown).
  • a human egg fluorescence assay was developed to characterize the binding of the various ZP antibodies to human zonae pellucidae.
  • the results of three different experiments are depicted in Fig. 6.
  • Serum against KLH and a normal mouse serum failed to give fluorescent staining whereas with all antisera against ZP components strong to intermediate levels of fluorescence were observed. From the presented data it can be inferred that antisera against ZP3(93-110), ZP3(172-190), ZP3(327-344), ZP3(341-360) and ZP3(362-372) recognize human oocytes.
  • the contraceptive potential of the ZP3 antibodies was analysed in a human sperm-zona binding assay. The results of this assay is presented in Fig. 7. Antibodies against total human zonae pellucidae showed 75% inhibition of sperm-zona binding. An antiserum against KLH does not interfere with sperm-egg binding. In contrast, a mixture of ZP3-peptide antibodies (peptides: ZP3(93-110), ZP3(172-190), ZP3(341-360) and ZP3(362-372)j showed a relative inhibition of sperm binding of 35%. Antibodies against recombinant ZP3 from CHO cells reduced sperm binding with 55%.
  • a 2.7 kb ZP3 'minigene' was assembled from the following three fragments: A, exon 1 and exon 2 joined in frame to each other by PCR-technology yielding a EcoRI-Xbal fragment: B, a genomic Xbal-Sall fragment with exon 3. 4 and part of exon 5; C. a Sall-Hindlll ZP3 cDNA fragment with exon 5 to 8 (provided with artificial 3' BamHI and Hindlll sites).
  • the three fragments were cloned in a pGEM vector (Promega) opened with EcoRI and Hindlll and subsequently as a 2.7 kb EcoRI-BamHI fragment placed behind the SV40 promoter in a mammalian expression vector.
  • control pEX; pEX-ZP3A KpnI-BamHI fragment; total cDNA
  • pEX-ZP3B KpnI-Sall fragment; cDNA truncated at exon 5 encoding amino acid 1-241
  • pEX-ZP3C KpnI-SphI fragment; cDNA truncated at end of exon 1 encoding amino acid 1-102
  • pEX-ZP3D Smal- BamHI fragment; 3' half of exon 5 + exon 6,7 and 8 encoding amino acid 257-372).

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Abstract

L'invention comprend un nouveau polypeptide ou un dérivé fonctionnel de ce dernier, possédant l'efficacité du ZP3 humain ou l'antigénicité du ZP3 humain. Elle comprend également des épitopes dudit polypeptide, des anticorps agissant contre les polypeptides ou les épitopes ainsi que des vaccins contraceptifs et des procédés d'expression des polypeptides dans un hôte approprié.
PCT/EP1991/001538 1990-08-27 1991-08-13 Proteine zp3 de la zona pellucida humaine Ceased WO1992003548A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
FI930897A FI930897A7 (fi) 1990-08-27 1991-08-13 Ihmisen munasolun keton proteiini ZP3
KR1019930700637A KR930702522A (ko) 1990-08-27 1991-08-13 인체의 투명층 단백질 제트피 3 (zp3)
NO93930718A NO930718L (no) 1990-08-27 1993-02-26 Zona pellucida menneske-protein zp3

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EP90202287 1990-08-27
EP90202287.0 1990-08-27

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WO1992003548A1 true WO1992003548A1 (fr) 1992-03-05

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PCT/EP1991/001538 Ceased WO1992003548A1 (fr) 1990-08-27 1991-08-13 Proteine zp3 de la zona pellucida humaine

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JP (1) JPH06500690A (fr)
KR (1) KR930702522A (fr)
CN (1) CN1060499A (fr)
AU (1) AU8328591A (fr)
CA (1) CA2090486A1 (fr)
FI (1) FI930897A7 (fr)
HU (1) HUT64394A (fr)
IE (1) IE912872A1 (fr)
NZ (1) NZ239518A (fr)
PT (1) PT98780A (fr)
WO (1) WO1992003548A1 (fr)
ZA (1) ZA916432B (fr)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994010304A1 (fr) * 1992-11-02 1994-05-11 Akzo Nobel N.V. Proteine zp3 de la zone pellucide du tamarin
WO1994011019A1 (fr) * 1992-11-09 1994-05-26 Zonagen, Inc. Materiaux et procedes utiles pour l'immunocontraception
WO1994022472A1 (fr) * 1993-03-26 1994-10-13 THE GOVERNMEMT OF THE UNITED STATES OF AMERICA as represented by THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES Vaccin contraceptif base sur l'allo-immunisation a l'aide de peptides de la zone pellucide
EP0558631A4 (en) * 1990-11-21 1995-11-22 Univ Washington Recombinant avirulent salmonella antifertility vaccines
WO1996006113A1 (fr) * 1994-08-22 1996-02-29 Akzo Nobel N.V. Nouveaux peptides immuno-contraceptifs
WO1997044358A1 (fr) * 1996-05-23 1997-11-27 Schering Aktiengesellschaft Proteines de membrane pellucide pour la contraception
WO1999042581A1 (fr) * 1998-02-19 1999-08-26 Eastern Virginia Medical School Proteine 3 (hzp3) humaine recombinee active de zona pellucida
US6001599A (en) * 1992-11-09 1999-12-14 Zonagen, Inc. DNAs encoding mammalian ZPBs
US7037663B2 (en) * 1998-02-19 2006-05-02 Eastern Virginia Medical School Human zona pellucida protein 3 and uses thereof
US7148021B2 (en) 2001-08-02 2006-12-12 Eastern Virginia Meical School Human zona pellucida proteins and methods of their use in diagnosing male infertility
CN101906163B (zh) * 2009-06-05 2012-06-27 上海交通大学医学院 一种免疫避孕的合成肽和嵌合肽及其应用
WO2020048924A1 (fr) * 2018-09-03 2020-03-12 Laboratoire Hra-Pharma Fragments de zp3 en immunothérapie du cancer de l'ovaire

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989003399A1 (fr) * 1987-10-07 1989-04-20 Zonagen, Inc. Procede de preparation et utilisation d'antigenes de zona pellucida et anticorps pour la sterilisation et la contraception
WO1990015624A1 (fr) * 1989-06-12 1990-12-27 The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce Vaccin contraceptif a base du gene de la zone pellucide clone

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989003399A1 (fr) * 1987-10-07 1989-04-20 Zonagen, Inc. Procede de preparation et utilisation d'antigenes de zona pellucida et anticorps pour la sterilisation et la contraception
WO1990015624A1 (fr) * 1989-06-12 1990-12-27 The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce Vaccin contraceptif a base du gene de la zone pellucide clone

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, vol. 110, no. 9, 1988, pages 425-426, abstract no. 73084e, (Columbus, Ohio, US), R.B. SHABANOWITZ et al.: "Molecular changes in the human zona pellucida associated with fertilization and human sperm-zona interactions", & ANN. N.Y. ACAD. SCI. 1988, 541, 621-32, see the whole abstract *
CHEMICAL ABSTRACTS, vol. 113, no. 11, 1990, pages 550-551, abstract no. 95872z, (Columbus, Ohio, US), R.B. SHABANOWITZ et al.: "Mouse antibodies to human zona pellucida: evidence that human ZP3 is strongly immunogenic and contains two distinct isomer chains", & BIOL. REPROD. 1990, 43(2), 260-70, see the whole abstract *
PROC. NATL. ACAD. SCI. USA, vol. 83, June 1986, pages 4341-4345; M.J. RINGUETTE et al.: "Oocyte-specific gene expression: Molecular characterization of a cDNA coding for ZP-3, the sperm receptor of the mouse zona pellucida" *
PROC. NATL. ACAD. SCI. USA, vol. 87, August 1990, M.E. CHAMBERLIN et al.: "Human homolog of the mouse sperm receptor", pages 6014-6018, see the whole article *

Cited By (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0558631A4 (en) * 1990-11-21 1995-11-22 Univ Washington Recombinant avirulent salmonella antifertility vaccines
WO1994010304A1 (fr) * 1992-11-02 1994-05-11 Akzo Nobel N.V. Proteine zp3 de la zone pellucide du tamarin
US5981228A (en) * 1992-11-09 1999-11-09 Zonagen, Inc. Mammalian ZPAs
US5989550A (en) * 1992-11-09 1999-11-23 Zonagen, Inc. Materials and methods for immunocontraception
US6027727A (en) * 1992-11-09 2000-02-22 Zonagen, Inc. Materials and methods for immunocontraception
AU675269B2 (en) * 1992-11-09 1997-01-30 Zonagen, Inc. Materials and methods for immunocontraception
US6001599A (en) * 1992-11-09 1999-12-14 Zonagen, Inc. DNAs encoding mammalian ZPBs
US5837497A (en) * 1992-11-09 1998-11-17 Zonagen, Inc. DNAs encoding mammalian ZPC and uses thereof
WO1994011019A1 (fr) * 1992-11-09 1994-05-26 Zonagen, Inc. Materiaux et procedes utiles pour l'immunocontraception
US5976545A (en) * 1992-11-09 1999-11-02 Zonagen, Inc. ZPC polypeptides
WO1994022472A1 (fr) * 1993-03-26 1994-10-13 THE GOVERNMEMT OF THE UNITED STATES OF AMERICA as represented by THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES Vaccin contraceptif base sur l'allo-immunisation a l'aide de peptides de la zone pellucide
EP1090641A3 (fr) * 1993-03-26 2002-06-05 THE GOVERNMENT OF THE UNITED STATES OF AMERICA as represented by the SECRETARY OF THE DEPARTMENT OF HEALTH AND HUMAN SERVICES Vaccin contraceptif base sur l'allo-immunisation a l'aide de peptides de la zone pellucide
WO1996006113A1 (fr) * 1994-08-22 1996-02-29 Akzo Nobel N.V. Nouveaux peptides immuno-contraceptifs
WO1997044358A1 (fr) * 1996-05-23 1997-11-27 Schering Aktiengesellschaft Proteines de membrane pellucide pour la contraception
AU723809B2 (en) * 1996-05-23 2000-09-07 Schering Aktiengesellschaft Zona pellucida proteins for contraception
US6344442B2 (en) * 1996-05-23 2002-02-05 Schering Aktiengesellschaft Zona pellucida proteins for contraception
WO1999042581A1 (fr) * 1998-02-19 1999-08-26 Eastern Virginia Medical School Proteine 3 (hzp3) humaine recombinee active de zona pellucida
US7019114B2 (en) * 1998-02-19 2006-03-28 Eastern Virginia Medical School Recombinant, biologically active human zona pellucida protein 3 (HZP3) to test male fertility
US7037663B2 (en) * 1998-02-19 2006-05-02 Eastern Virginia Medical School Human zona pellucida protein 3 and uses thereof
US7148021B2 (en) 2001-08-02 2006-12-12 Eastern Virginia Meical School Human zona pellucida proteins and methods of their use in diagnosing male infertility
CN101906163B (zh) * 2009-06-05 2012-06-27 上海交通大学医学院 一种免疫避孕的合成肽和嵌合肽及其应用
WO2020048924A1 (fr) * 2018-09-03 2020-03-12 Laboratoire Hra-Pharma Fragments de zp3 en immunothérapie du cancer de l'ovaire

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HU9300537D0 (en) 1993-05-28
IE912872A1 (en) 1992-03-11
JPH06500690A (ja) 1994-01-27
FI930897A0 (fi) 1993-02-26
KR930702522A (ko) 1993-09-09
FI930897L (fi) 1993-04-05
CN1060499A (zh) 1992-04-22
FI930897A7 (fi) 1993-04-05
ZA916432B (en) 1992-06-24
PT98780A (pt) 1992-07-31
NZ239518A (en) 1994-03-25
HUT64394A (en) 1993-12-28
CA2090486A1 (fr) 1992-02-28
AU8328591A (en) 1992-03-17

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